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Transcript
Polymerase Chain Reaction ⢠Based on DNA polymerase, the enzyme that replicates DNA. â Needs template DNA and two primers that flank the region to be amplified. Primers are short (generally 18-30 bases) DNA oligonucleotides complementary to the ends of the region being amplified. â Starting at each primer, DNA polymerase adds new bases to the 3' ends to create the new second strand. â PCR is a cyclical process. Each cycle doubles the number of DNA molecules between the primers: exponential growth. â A key element in PCR is a special form of DNA polymerase from Thermus aquaticus, a bacterium that lives in nearly boiling water in the Yellowstone National Park hot springs. This enzyme, Taq polymerase, can withstand the temperature cycle of PCR, which would kill DNA polymerase from E. coli. ⢠advantages: â rapid, sensitive, lots of useful variations, robust (works even with partly degraded DNA) ⢠disadvantages: â Only short regions (up to 2 kbp) can be amplified. â limited amount of product made
